LOCUS       sequence-315863-        3942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: GCaMP6F calcium sensor (cytosolic).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3942)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 3942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS008E: Entry vector for: GCaMP6F calcium se...
FEATURES             Location/Qualifiers
     source          1..3942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      674..683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(680..688,689..706,707..709,710..742,743..745,746..769,
                     770..796,797..853,854..859,860..1129,1130..1147,1148..1582,
                     1583..1588,1589..2029)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      776..785
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1066..1087
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1193..1214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1454..1473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2067..2166)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2183..2202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2184..2202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2207..2223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2207..2223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2325..2344)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2336..3145
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2403..2422)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3292..3880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3781..3800
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctga tccgccacca tgggttctca tcatcatcat catcatggta tggctagcat
      721 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atctcgccac
      781 catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca gagctatagg
      841 tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga acggcatcaa
      901 ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta
      961 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1021 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1081 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     1141 agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1201 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     1261 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1321 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1381 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     1441 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1501 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1561 ggggcacaag ctggagtaca acctgccgga ccaactgact gaagagcaga tcgcagaatt
     1621 taaagaggaa ttctccctat ttgacaagga cggggatggg acaataacaa ccaaggagct
     1681 ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat
     1741 caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat
     1801 ggcaagaaaa atgaaataca gggacacgga agaagaaatt agagaagcgt tcggtgtgtt
     1861 tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga tgacaaacct
     1921 tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag acatcgatgg
     1981 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt acaagttctg
     2041 ctacgagaac gaggtgtaag aattcgaccc agctttcttg tacaaagttg gcattataag
     2101 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2161 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2221 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2281 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2341 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2401 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2461 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2521 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2581 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2641 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2701 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2761 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2821 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2881 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2941 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3001 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3061 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3121 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3181 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3241 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3901 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
